陆地棉原生质体培养与植株再生

Protoplast Culture and Plant Regeneration in Upland Cotton

以陆地棉品种“珂字２０１”为材料，比较了ＩＡＡ＋ＫＴ和２，４－Ｄ＋ＫＴ在愈伤诱导和悬浮培养中的效应，结果表明，愈伤组织诱导中，ＩＡＡ和２，４－Ｄ表现为正效应，且２，４－Ｄ的效应强于ＩＡＡ；ＫＴ表现为负效应；胚性愈伤悬浮培养中，３种激素都表现出负效应。以胚性细胞悬浮系为材料进行了原生质体的分离和培养试验，分离原生质体的最佳酶组合为纤维素酶（ＯｎｏｚｕｋａＲ１０）３％＋果胶酶（Ｐｅｃｔｉｎａｓｅ）１．５％，原生质体培养的最佳激素组合为ＩＡＡ０．５ｍｇ／Ｌ＋ＫＴ０．１ｍｇ／Ｌ，适当提高原生质体密度有利于原生质体培养。光照，温度等物理因素对原生质体培养也有明显影响。原生质体再生的愈伤组织直接在无激素的悬浮培养基中增殖，很快获得了大量胚性愈伤组织。

In callus induction of Gossypium hirsutum L.cv.coker 201,IAA and 2,4 D showed positive effect and KT showed negative effect,while negative effect was shown with the three hormones in suspensions.The best enzymatic combination in protoplast isolation from embryogenetic suspensions was 3% cellulase(Onozuka R10)+1.5%pectinase,and the best hormone combination in protoplast culture was IAA 0.5mg/L+KT 0.1mg/L. Appropriately increasing protoplast density benefited protoplast culture. Light and temprature affected the protoplast culture.Protoplast derived calli growed rapidly in suspension culture medium without any hormone and regenerated plants through differentiation culture.